Abstract - We characterized the mitochondrial DNA of two stingless bee species of the genus Partamona. Partial restriction maps were obtained based on digestion of PCR amplified fragments with 8 restriction enzymes. Using Melipona bicolor mtDNA sequence as a model, we were able to amplify 12120 bp of P. mulata and 10300 bp of P. helleri, about 65.5% and 55.7% of their mitochondrial genome, respectively. The digestion assays showed 16 restriction sites for P. mulata and 20 for P. helleri, some of which were exclusive to the genus and others shared with other Meliponini species. The main mitochondrial genes could be mapped and through sequencing analysis we verified that the intergenic region that occurs between the genes CO I and CO II in Apis is absent in Partamona.